Fig. 6

ABI5 functions genetically downstream of ECT2/ECT3/ECT4. a Relative mRNA levels of ABI5 in 7-day-old WT and ect2/3/4 seedlings under Mock and ABA treatment. TUB8 was used as the internal control gene. Data are presented as means ± SE, n = 3 biological replicates × 2 technical replicates. *P < 0.05, **P < 0.01 (two-sided t-test). b Western blotting showing the ABI5 protein level in 7-day-old WT and ect2/3/4 seedlings under ABA treatment. The relative abundance of ABI5 in WT was set to 1 by normalized to the loading control (β-Actin2). c Phenotypic analysis of the ABA response in WT, ect2/3/4, abi5-10, and Crispr ABI5/ect2/3/4 seeds grown on 1/2 MS-medium supplemented with 0 (Mock) and 0.5 μM ABA under long-day conditions. Representative photographs were taken 8 days after cold stratification. d Statistical analysis of germination rates in WT, ect2/3/4, abi5-10, and Crispr ABI5/ect2/3/4 plants under ABA treatment. At least 30 seeds per genotype were measured in each replicate. Biological triplicates were averaged. Data are presented as the mean ± SE. e Statistical analysis of germination rates 4 days after imbibition and of cotyledon greening rates 8 days after imbibition in WT, ect2/3/4, abi5-10, and Crispr ABI5/ect2/3/4 plants under ABA treatment. Data are presented as the mean ± SE; n = 3 biological replicates. ***P < 0.001, ****P < 0.0001 compared with WT (two-sided t-test). f Relative mRNA expression levels of RD29A and EM6 in 7-day-old WT, ect2/3/4, abi5-10, and Crispr ABI5/ect2/3/4 seedlings under Mock and ABA treatment. TUB8 was used as the internal control gene. Data are presented as means ± SE, n = 2 biological replicates × 2 technical replicates. *P < 0.05 compare with WT (two-sided t-test)